Enhancement of Labneh Quality by Laser-Induced Modulation of Lactocaseibacillus casei NRRL B-1922

Round 1

Reviewer 1 Report

1. It is necessary to unify all the names of microorganisms in the publication;
2. Despite the fact that the authors cite their development for laser measurement, it is good to show the main positions of the measurement in order to make the description of the method clearer.
3. Recommends that the description of the experiment be supplemented by a graphical representation (item 2.6), as it currently sounds vague.
4. On page 6, lines 214 to 217, it is claimed that there is a significant reduction in the number of viable cells, but this is not fully supported by Figure 2. Please correct!
5. Item 3.4 states that there is a significant increase in the DPPH indicator. Please correct, as the results do not fully support the statement!
6. In item 3.5, please reconsider the statement "significantly"!

Author Response

Firstly, we would like to express our thanks to the reviewer, for his precious time and constructive suggestions. These contributions made the manuscript clearer to the reader and improved its quality.

We ‎have carefully read all comments and suggestions and made as much modifications as we ‎believed possible to meet the reviewers’ requests. Questions are individually presented and ‎immediately answered. The changes are yellow highlighted in the revised manuscript.

 

1. It is necessary to unify all the names of microorganisms in the publication;

Thanks for your suggestion, the names of Lactobacillus casei and Lactobacillus bulgaricus were unified to the new name Lacticaseibacillus casei and Lactobacillus delbrueckii spp. bulgaricus respectively.

1. Despite the fact that the authors cite their development for laser measurement, it is good to show the main positions of the measurement in order to make the description of the method clearer.

Thanks for your suggestion, a new sentence was added to clarify the method “line 139-140”

1. Recommends that the description of the experiment be supplemented by a graphical representation (item 2.6), as it currently sounds vague.

The reviewer suggestion is appreciated, and a new graphical representation was inserted in the manuscript as figure 2

1. On page 6, lines 214 to 217, it is claimed that there is a significant reduction in the number of viable cells, but this is not fully supported by Figure 2. Please correct!

The bacterial viable counts were decreased after cold storage, but that decrease was not significant. Such findings indicated the stability of all bacterial culture during cold storage and the laser treatments did not have any negative effect on the viability of L. casei. The sentence was revised “line 217-218” page 6 and “line 305” page 10.

1. Item 3.4 states that there is a significant increase in the DPPH indicator. Please correct, as the results do not fully support the statement!

In this part, we compared the antioxidant activities at the beginning of cold storage (Day 1) between the different experimental groups. The results were significantly different (control; 40.56±1.20 f, 3 J/cm²; 45.61±1.35 de, 6 J/cm²; 50.78±2.01 d ,12 J/cm²; 56.39±0.92 c). The same was done after 20 days storage. We did not mean significant differences between control (before and after cold storage) and treatment (before and after cold storage). We modified the sentence to be clearer “Line 228 page 7”

1. In item 3.5, please reconsider the statement "significantly"!

The sentence was modified and clarified, since the observed significant difference was only after 20 days of cold storage when compared with control with the same storage period. We changed the table representation for better expression and easily understood by the reader “Line 236 page 8”.

Author Response File: Author Response.docx

Reviewer 2 Report

A very interesting study on the production of Labneh. The use of a laser to irradiate high-dose bacteria before milk fermentation seems to be an innovative method, positively influencing the properties of the final product. In the opinion of the reviewer, the research was planned correctly and the research methods did not raise any objections. The research methods were properly described. The research results were collected in the form of tables and graphs. The obtained results were supported by statistical analysis, which allows for their more precise interpretation. At the same time, it should be noted that the obtained results are promising and offer the possibility of translating them into other strains of lactic acid bacteria. The summary includes the most important elements of the work, they are clearly and concisely written. The literature has been cited correctly and adequately to the topic.  In the opinion of the reviewer, I suggest accepting the manuscript for publication

Author Response

All authors would like to thank Reviewer#2 for his valuable comments and positive feedback. The reviewer’s words have a deep impact on us as they stimulated our motivation and passion towards our research work. We express our sincere appreciation and gratitude to the reviewer.

 

Author Response File: Author Response.docx

Reviewer 3 Report

The manuscript ‘Enhancement of Labneh quality by Laser induced modulation of Lactocaseibacillus casei NRRL B-1922” is a trial to study the effect of He-Ne laser with different doses on the biomodulation of L. casei used in the milk fermentation for Labneh production. The scope is an interesting for potential reader in a surrounding needs for probiotic product. The paper is readable but needs some minor improvement before publication in Fermentation journal.

Minor comments:

l. 36 – The justification of such improvements (41 and 14%) in the manuscript should be clearly stated

l. 37 – the explanation of the ‘significant improved biological effects on the final products’ should be clearly stated

l. 183 – explanation if panelists were trained is needed, and the description of the scale is needed

l. 302 – Figure 3 is missing

Author Response

Firstly, we would like to express our thanks to the reviewer, for his precious time and constructive suggestions. These contributions made the manuscript clearer to the reader and improved its quality.

We ‎have carefully read all comments and suggestions and made as much modifications as we ‎believed possible to meet the reviewers’ requests. Questions are individually presented and ‎immediately answered. The changes are yellow highlighted in the revised manuscript.

 

Minor comments:

 

1. 36 – The justification of such improvements (41 and 14%) in the manuscript should be clearly stated

 

Thanks for the suggestion. Despite we have mentioned clearly the justification of the antioxidant capacity improvement in the manuscript at discussion section Line 312-321, we have added new sentences highlighted in yellow to follow the reviewer’s valuable suggestion.

 

1. 37 – the explanation of the ‘significant improved biological effects on the final products’ should be clearly stated

Per the reviewer’s valuable remark and advice, we have changed it to “significant improved selected characteristics on the final products”. The selected characteristics including flavor aiding components, the antioxidant and proteolytic activities). They were explained in detail just before this sentence of the abstract.

 

1. 183 – explanation if panelists were trained is needed, and the description of the scale is needed

We modified the method to be more detailed “line 190-191” to show that the participated panelists were adequately trained.

 

1. 302 – Figure 3 is missing

We have corrected this typo error and thanks to the great observation from the reviewer.

Author Response File: Author Response.docx

Round 2

Reviewer 1 Report

The authors have taken into account all recomendations. 

This manuscript is a resubmission of an earlier submission. The following is a list of the peer review reports and author responses from that submission.

Round 1

Reviewer 1 Report

1. In the discussion, the research data in the table should also be clearly stated, not only referring to the reading of the table.
2. the cited references are currently more than 5 years (there are still references from the 1990s)
3. please explain the mechanism of the effect of laser-induced on each of the tests carried out (see the comment in the article)

Author Response

Point by point Response to Reviewer#1

Firstly, we would like to express our thanks to the reviewer, for your time and constructive suggestions. These contributions made the manuscript clearer to the reader and improved its quality.

We ‎have read carefully all comments and suggestions and made as much modifications as we ‎believed possible to meet the reviewers’ requests. Questions are individually presented and ‎immediately answered. The changes are highlighted in yellow in the revised manuscript.

1. In the discussion, the research data in the table should also be clearly stated, not only referring to the reading of the table.

Thanks for your suggestion, the discussion was thoroughly rewritten and revised. Please find the modified discussion “line 240-322 pages 10,11,12”.

 

2. the cited references are currently more than 5 years (there are still references from the 1990s)

Thanks for your suggestion and your valuable comment. We have inserted new and updated references. We have as well updated other references making more than 25% of the used references from the last two years only.

3. please explain the mechanism of the effect of laser-induced on each of the tests carried out (see the comment in the article)

We have explained and discussed the effects of laser induction in the rewritten discussion section of the manuscript per the reviewer’s valuable comment.

We also referred the reader to our explanation in our previous work. In Brief; “Despite that the exact action mechanism of PBM is not yet well understood, low laser doses can have altered actions by cellular signaling induction in different routes. For microorganisms like Escherichia coli, cytochrome bd and bo3 complexes are found to be the key player in the biological alterations induced by light absorption. They are responsible for stimulating transduction processes which lead to intracellular signaling pathways, and hence enlarging the effect of the primary absorbed light signal. The molecules that are recruited in transducing such signals are chemically reactive species as reactive oxygen, whereby they react with other biomolecules and cellular structures to change their functions and/or expressed gene.”

 

Author Response File: Author Response.docx

Reviewer 2 Report

General comments:

1. The topic of this paper is relevant, timely, and of interest to the audience of this journal but there are some inquiries and comments are mentioned below, therefore the manuscript is not acceptable for publication in this form.
2. A native speaker or somebody good in English should revise the In many sections meaning is lost due to poor English. See below corrections that can improve the manuscript.

Specific comments:

In title:

1. I suggest that the title modify to “Enhancement of Labneh quality by Laser induced modulation of Lactobacillus casei NRRL B-1922”.
2. How do you know this LAB strain (Lactocaseibacillus casei B-1922) is a probiotic bacterium? the site of NRRL laboratory did not mention that??!!

In abstract section:

1. In line 20: please change “Labneh products” to Labneh product” without “s” and through the manuscript.
2. In line 20: change “are” to “is”, delete “nowadays”
3. In line 21: change “worldwide” to “all over the world”
4. In line 23: change “affects” to “affect”
5. In line 27: change “enhanced” to “enhancing”
6. In lines 33-37: “Moreover, the study investigated ……….” should be re-

In introduction section:

1. In lines 44-45: definition of Labneh should be re-phrased
2. In line 45: delete “in in” to become “in”
3. In line 49: add “of Labneh” after “the industrial process”
4. In line 54: change “byproduct” to “metabolite”
5. In line 57: change “marked” to “market”
6. In line 61: add “full” before “fat Labneh” to become “as full fat Labneh”
7. In line 65: add “lactic acid bacteria” after “thermophilic” and delete “where they are employed”
8. In lines 66-67: the name of microorganisms should be written in italic and through the manuscript such as in lines 82, 83, 84, and etc.
9. In line 68: change “shortens” to “reduces, decreases or declines”
10. In line 71: change “yoghurt products” to “yoghurt products”
11. In line 72: add coma “ , “ after “as the former” then add “it may”
12. In line 74: delete “Yet”
13. In line 76: modify “the minor reduction of the viable bacterial count of Labneh by ….”
14. In lines 80-81: “after surviving the hosts gastrointestinal tract” not clear.
15. In line 81: “that’s why they are ………” what are the causes to use probiotic bacteria? this is not clear in the last sentence!!
16. In line 85: what does mean of “yeast and bacilli strains”? does mean Lactobacilli or molds and yeasts??!!
17. In line 94: what about “UV-B”???
18. In line 102: put “on the other hand” at the first of sentence then add “ , “
19. In line 103: what does mean of “ischemic damage related diseases”

In Material and methods section:

1. In line 114: please, add the other chemicals and microbiological media.
2. In line 132: change “10” to “Ten”
3. In line 149: re-phrase a sentence to “either fresh (day 1) and after twenty days of cold storage”
4. In line 152: should be revised the format of journal in references style in text where should be excluded “the year” if the references were mentioned numerical through the manuscript.
5. In line 154: delete “Development of flavor components”
6. In lines 164-165: re-phrase a sentence to “The DPPH scavenging radical activity of different labneh treatments was assessed as described by …….”   
7. In line 168: How did prepare the crude extract to determine the proteolytic and antioxidant activity? It should be mentioned.
8. In line 170: “o” should be written in capital and italic “O” and change “whey” to “crude extract”
9. In line 172: “wasmeasured” to “was measured”
10. In line 184: (SAS, 1994) not found in the references list as a reference??!!

In results section:

1. In line 188: it should be illustrated the results at day 1 of manufacturing of labneh.
2. In line 206: start with “after 20 days of cold storage” not “during”
3. In Table 3: Why was the result of ascorbic acid as positive control not included in the statistical analysis???
4. Under proteolytic activity: in my opinion, presence of B. subtilis in labneh treatments affected on the proteolytic activity certainly, particularly the viable count of B. subtilis were increased after storage period. How did you overcome or explain that???

In discussion section:

The discussion is very poor and needs for focusing on the analysis of the obtained results, their interpretations, and their relevance for the advancement of knowledge about the potential use of photo-biomodulation or induction with Laser beam.

In references list:

The references are not in the right format according to the style of journal. Authors should be revised the format.

Author Response

Point by point Response to Reviewer#2

Firstly, we would like to express our deep appreciation and thanks to the editor and the reviewer for their sincere advices and valuable remarks and notes. These contributions made the manuscript clearer to the reader and improved its quality.

We ‎have read carefully all comments and suggestions and made as much modifications as we ‎believed possible to meet the reviewers’ requests. Questions are individually presented and ‎immediately answered. The changes are highlighted in yellow in the revised manuscript.

 

1. The topic of this paper is relevant, timely, and of interest to the audience of this journal but there are some inquiries and comments are mentioned below, therefore the manuscript is not acceptable for publication in this form.
2. A native speaker or somebody good in English should revise the In many sections meaning is lost due to poor English. See below corrections that can improve the manuscript.

The Manuscript has been revised

Specific comments:

In title:

1. I suggest that the title modify to “Enhancement of Labneh quality by Laser induced modulation of Lactobacillus casei NRRL B-1922”.

Thanks for your constructive suggestion, the title was modified

“Lacticaseibacillus casei formerly named as Lactobacillus casei are reclassified recently based on the whole genome sequences analysis to novel genera; Lacticaseibacillus (the L. casei group) (Zheng et al. 2020). This new genus remains relatively heterogeneous and the strains belonging to this genus have a considerable economic importance as many species that are used as starter cultures in dairy fermentations as well as probiotics (Zheng et al. 2020).”

 

2. How do you know this LAB strain (Lactocaseibacillus casei B-1922) is a probiotic bacterium? the site of NRRL laboratory did not mention that??!!

There are many studies demonstrated the probiotic properties of the strain Lactocaseibacillus casei B-1922 (for example; Mani-López et al. 2014; doi:10.3168/jds.2013-7551).

In addition, we previously characterized some of the probiotic properties of this strain (Mohamed et al., 2020; doi:10.3390/biology9090256) 

In abstract section:

1. In line 20: please change “Labneh products” to Labneh product” without “s” and through the manuscript.

Done

2. In line 20: change “are” to “is”, delete “nowadays”

Done

3. In line 21: change “worldwide” to “all over the world”

Done

4. In line 23: change “affects” to “affect”

Done

5. In line 27: change “enhanced” to “enhancing”

Done

6. In lines 33-37: “Moreover, the study investigated ……….” should be re-phrased

Done, the sentence was re-phrased, please see line 33-35

In introduction section:

1. In lines 44-45: definition of Labneh should be re-phrased

Done, the sentence was re-phrased, please see line 42-43

2. In line 45: delete “in in” to become “in”

Done

3. In line 49: add “of Labneh” after “the industrial process”

Done

4. In line 54: change “byproduct” to “metabolite”

Done

5. In line 57: change “marked” to “market”

Done

6. In line 61: add “full” before “fat Labneh” to become “as full fat Labneh”

Done

7. In line 65: add “lactic acid bacteria” after “thermophilic” and delete “where they are employed”

Done

8. In lines 66-67: the name of microorganisms should be written in italic and through the manuscript such as in lines 82, 83, 84, and etc.

Done, the manuscript was thoroughly revised.

9. In line 68: change “shortens” to “reduces, decreases or declines”

Done

10. In line 71: change “yoghurt products” to “yoghurt products”

Done

11. In line 72: add coma “ , “ after “as the former” then add “it may”

Done

12. In line 74: delete “Yet”

Done

13. In line 76: modify “the minor reduction of the viable bacterial count of Labneh by ….”

Done

14. In lines 80-81: “after surviving the hosts gastrointestinal tract” not clear.

The paragraph was rephrased please see line 78-80

15. In line 81: “that’s why they are ………” what are the causes to use probiotic bacteria? this is not clear in the last sentence!!

The last sentence is rephrased “line78-80”

16. In line 85: what does mean of “yeast and bacilli strains”? does mean Lactobacilli or molds and yeasts??!!

Corrected to be clearer into “some molds and yeast strains”

17. In line 94: what about “UV-B”???

Done

18. In line 102: put “on the other hand” at the first of sentence then add “ , “

Done

19. In line 103: what does mean of “ischemic damage related diseases”

Done

In Material and methods section:

1. In line 114: please, add the other chemicals and microbiological media.

Done

2. In line 132: change “10” to “Ten”

Done

3. In line 149: re-phrase a sentence to “either fresh (day 1) and after twenty days of cold storage”

Done, corrected

4. In line 152: should be revised the format of journal in references style in text where should be excluded “the year” if the references were mentioned numerical through the manuscript.

Done, these have been corrected.

5. In line 154: delete “Development of flavor components”

Done

6. In lines 164-165: re-phrase a sentence to “The DPPH scavenging radical activity of different labneh treatments was assessed as described by …….”

Done, please see Line 170-172

7. In line 168: How did prepare the crude extract to determine the proteolytic and antioxidant activity? It should be mentioned.

Done please see Line 169-170

8. In line 170: “o” should be written in capital and italic “O” and change “whey” to “crude extract”

Done

9. In line 172: “was measured” to “was measured”

Done

10. In line 184: (SAS, 1994) not found in the references list as a reference??!!

Done

 

In results section:

1. In line 188: it should be illustrated the results at day 1 of manufacturing of labneh.

Done

2. In line 206: start with “after 20 days of cold storage” not “during”

Done

3. In Table 3: Why was the result of ascorbic acid as positive control not included in the statistical analysis???

Done, the ascorbic acid is included in the statistical analysis

4. Under proteolytic activity: in my opinion, presence of subtilis in labneh treatments affected on the proteolytic activity certainly, particularly the viable count of B. subtilis were increased after storage period. How did you overcome or explain that???

Per the reviewer’s valuable remark and advise, the aim of labneh spoilage experiment by B. subtilis and E. coli during cold storage was to evaluate the viable count number of spoilage bacteria in control and laser treatment. The results indicate significant increase in number of B. subtilis in control sample (without treatment) however the increase in the number was not significant in the laser treatment although the laser treatment increase the proteolytic activities, this could be attributed to the antibacterial effects of L. casei secondary metabolites after pre-treatment with red laser through the high concentration of organic acids or via the enhanced production of antimicrobial peptides (Mohamed et al., 2020). Therefore, it can be concluded that L. casei inhibit B. subtilis growth during shelf-life storage.

In discussion section:

 

The discussion is very poor and needs for focusing on the analysis of the obtained results, their interpretations, and their relevance for the advancement of knowledge about the potential use of photo-biomodulation or induction with Laser beam.

Thanks for your suggestion and comments. Most of the discussion part was rewritten and new references inserted, and we did our best to clarify all the mentioned points. Please find the modified discussion “line 240-322 pages 10,11,12”.

In references list:

The references are not in the right format according to the style of journal. Authors should be revised the format.

Done, corrected to the style of journal.

Author Response File: Author Response.docx

Round 2

Reviewer 2 Report

I believe the manuscript was sufficiently improved to warrant publication in Fermentation journal.